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. 2022 Feb 16;12:819554. doi: 10.3389/fcimb.2022.819554

Figure 6.

Figure 6

CFTR modulators restore defective autophagy in F508del macrophages. (A) Representative LC3 immunoblot from wild-type (WT) and F508del macrophages treated with either vehicle (non-infected (NT)) or 10 µM of Teza −/+ 5 µM of Iva for 24 h, followed by −/+100 nM of Baf-A1 (n = 4 biological replicates). (B) Densitometry analysis for LC3II in WT and F508del macrophages shown (A), representing the ratio of LC3II to GAPDH. Data represent mean ± SEM (n = 5 WT, n = 7 cystic fibrosis (CF)). Statistical analysis was performed using two-way ANOVA; *, p ≤ 0.05; **, p ≤ 0.01; ns, non-significant. (C) Confocal microscopy images showing LC3 puncta accumulation in F508del macrophages either NT or treated with Teza+Iva −/+ Baf-A1 (100 nM) (n = 3 biological replicates). Scale bar = 10 µm. (D) Percentage of cells expressing >5 LC3 puncta in F508del macrophages shown in panel (C) Data represent mean ± SEM calculated by scoring at least 200 cells from randomly chosen fields of view, normalized to the total number of cells from 3 independent experiments. Statistical analysis was performed using paired t-test; *, p ≤ 0.05. (E) LC3 integrated density in F508del macrophages either NT or Teza+Iva treated −/+ Baf-A1. Data represent mean ± SEM calculated using ImageJ software from at least 5 randomly chosen fields of view with approximately 200 cells from 5 independent experiments. Statistical analysis was performed using one-way ANOVA; *, p ≤ 0.05; ns, non-significant.